Abstract The antimicrobial substance triclosan has widespread use in personal care products and can enter the terrestrial environment if sewage sludge is applied to soil. The inhibitory effects of triclosan on basal and substrate-induced respiration (SIR) of three different soils were investigated. Soils were dosed and later redosed with four nominal triclosan concentrations, and respiration rates were measured over time. In each soil, a significant depression in basal respiration was noted after initial dosing, followed by a recovery. The initial extent of respiration inhibition was positively related to the triclosan dose, i.e., respiration was most inhibited at highest triclosan concentration. Differences in respiration inhibition between soils at equivalent dose were inversely correlated with organic matter and clay content, suggesting that the bioavailability of triclosan might have been reduced by sorption to organic carbon or by physical protection in micropores. Substrate-induced respiration was also reduced by the addition of triclosan and subsequently recovered. After redosing with triclosan, basal respiration was enhanced in all soils, suggesting that it was acting as a substrate. However, redosing resulted in SIR inhibition in all treatments above 10 mg triclosan kg−1 in all three soils, although all soils appeared to be more resistant to perturbation than following initial dosing. The present study suggests that triclosan inhibits soil respiration but that a subsequent acclimation of the microbial community occurs. Environ. Toxicol. Chem. 2011;30:360–366. © 2010 SETAC